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1.
Chemistry ; 30(16): e202302924, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38242847

RESUMEN

Two Mo(0) phosphenium complexes containing ancillary secondary phosphine ligands have been investigated with respect to their ability to participate in electrophilic addition at unsaturated substrates and subsequent P-H hydride transfer to "quench" the resulting carbocations. These studies provide stoichiometric "proof of concept" for a proposed new metal-catalyzed electrophilic hydrophosphination mechanism. The more strongly Lewis acidic phosphenium complex, [Mo(CO)4(PR2H)(PR2)]+ (R=Ph, Tolp), cleanly hydrophosphinates 1,1-diphenylethylene, benzophenone, and ethylene, while other substrates react rapidly to give products resulting from competing electrophilic processes. A less Lewis acidic complex, [Mo(CO)3(PR2H)2(PR2)]+, generally reacts more slowly but participates in clean hydrophosphination of a wider range of unsaturated substrates, including styrene, indene, 1-hexene, and cyclohexanone, in addition to 1,1-diphenylethylene, benzophenone, and ethylene. Mechanistic studies are described, including stoichiometric control reactions and computational and kinetic analyses, which probe whether the observed P-H addition actually does occur by the proposed electrophilic mechanism, and whether hydridic P-H transfer in this system is intra- or intermolecular. Preliminary reactivity studies indicate challenges that must be addressed to exploit these promising results in catalysis.

2.
Blood Purif ; 52(4): 366-372, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36702111

RESUMEN

INTRODUCTION: Exit-site infection (ESi) prevention is a key factor in lowering the risk of peritonitis. This study aimed to evaluate the associations between exit-site (ES) care protocols and the annual incidence rates of ESi and peritonitis in Portugal. METHODS: We performed a national survey using two questionnaires: one about the incidence of catheter-related infections and the other characterizing patients' education and ES care protocols. RESULTS: In 2017 and 2018, 14 Portuguese units followed 764 and 689 patients. ESi incidence rate was 0.41 episodes/year, and the peritonitis incidence rate was 0.37. All units monitor catheter-related infections on a yearly basis, use antibiotic prophylaxis at the time of catheter placement, and treat nasal carriage of S. aureus, although with different approaches. Screening for nasal carriage of S. aureus is performed by 12 units, and daily topical antibiotic cream is recommended by 6 out of 14 of the units. We did not find statistical differences in ESi/peritonitis, comparing these practices. The rate of ESis was lower with nonocclusive dressing immediately after catheter insertion, bathing without ES dressing, with the use of colostomy bags in beach baths and was higher with the use of bath sponge. The peritonitis rate was lower with bathing without ES dressing and if shaving of the external cuff was performed in the presence of chronic ESi. CONCLUSIONS: We found potential proceedings associated with ESi and peritonitis. A regular national audit of peritoneal dialysis units is an important tool for clarifying the best procedures for reduction of catheter-related infections.


Asunto(s)
Infecciones Relacionadas con Catéteres , Diálisis Peritoneal , Peritonitis , Humanos , Mupirocina , Portugal , Infecciones Relacionadas con Catéteres/etiología , Staphylococcus aureus , Catéteres de Permanencia/efectos adversos , Administración Tópica , Diálisis Renal/efectos adversos , Antibacterianos , Diálisis Peritoneal/efectos adversos , Peritonitis/etiología
3.
Mol Biol Evol ; 33(1): 93-108, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26420451

RESUMEN

The human kallikrein (KLK) cluster, located at chromosome 19q13.3-13.4, encodes 15 serine proteases, including neighboring genes (KLK3, KLK2, KLK4, and KLK5) with key roles in the cascades of semen liquefaction, tooth enamel maturation, and skin desquamation. KLK2 and KLK3 were previously identified as targets of adaptive evolution in primates through different mechanisms linked to reproductive biology and, in humans, genome-wide scans of positive selection captured, a yet unexplored, evidence for KLK neutrality departure in East Asians. We perform a detailed evaluation of KLK3-KLK5 variability in the 1000 Genomes samples from East Asia, Europe, and Africa, which was sustained by our own sequencing. In East Asians, we singled out a 70-kb region surrounding KLK4 that combined unusual low levels of diversity, high frequency variants with significant levels of population differentiation (FST > 0.5) and fairly homogenous haplotypes given the large local recombination rates. Among these variants, rs1654556_G, rs198968_T, and rs17800874_A stand out for their location on putative regulatory regions and predicted functional effects, namely the introduction of several microRNA binding sites and a repressor motif. Our functional assays carried out in different cellular models showed that rs198968_T and rs17800874_A operate synergistically to reduce KLK4 expression and could be further assisted by rs1654556_G. Considering the previous findings that KLK4 inactivation causes enamel malformations in humans and mice, and that this gene is coexpressed in epidermal layers along with several substrates involved in either cell adhesion or keratinocyte differentiation, we propose KLK4 as another target of selection in East Asians correlated to tooth and epidermal morphological traits.


Asunto(s)
Pueblo Asiatico/genética , Regulación hacia Abajo/genética , Variación Genética/genética , Calicreínas/genética , Evolución Molecular , Asia Oriental , Humanos , Selección Genética/genética
4.
J Lipid Res ; 56(11): 2158-71, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26351365

RESUMEN

Cytotoxic bile acids, such as deoxycholic acid (DCA), are responsible for hepatocyte cell death during intrahepatic cholestasis. The mechanisms responsible for this effect are unclear, and recent studies conflict, pointing to either a modulation of plasma membrane structure or mitochondrial-mediated toxicity through perturbation of mitochondrial outer membrane (MOM) properties. We conducted a comprehensive comparative study of the impact of cytotoxic and cytoprotective bile acids on the membrane structure of different cellular compartments. We show that DCA increases the plasma membrane fluidity of hepatocytes to a minor extent, and that this effect is not correlated with the incidence of apoptosis. Additionally, plasma membrane fluidity recovers to normal values over time suggesting the presence of cellular compensatory mechanisms for this perturbation. Colocalization experiments in living cells confirmed the presence of bile acids within mitochondrial membranes. Experiments with active isolated mitochondria revealed that physiologically active concentrations of DCA change MOM order in a concentration- and time-dependent manner, and that these changes preceded the mitochondrial permeability transition. Importantly, these effects are not observed on liposomes mimicking MOM lipid composition, suggesting that DCA apoptotic activity depends on features of mitochondrial membranes that are absent in protein-free mimetic liposomes, such as the double-membrane structure, lipid asymmetry, or mitochondrial protein environment. In contrast, the mechanism of action of cytoprotective bile acids is likely not associated with changes in cellular membrane structure.


Asunto(s)
Apoptosis , Ácido Desoxicólico/farmacología , Membranas Mitocondriales/metabolismo , Animales , Supervivencia Celular , Células Cultivadas , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Masculino , Fluidez de la Membrana , Mitocondrias Hepáticas/efectos de los fármacos , Mitocondrias Hepáticas/metabolismo , Permeabilidad , Ratas , Transducción de Señal
5.
Artif Organs ; 39(2): 142-9, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25277688

RESUMEN

Hemodiafiltration (HDF) with 20-22 L of substitution fluid is increasingly recognized as associated with significant benefits regarding patient outcome. However, some doubt exists as to whether these high volumes can be achieved in routine clinical practice. A total of 4176 sessions with 366 patients on postdilution HDF were analyzed in this 1-month observational cohort study with prospective data collection. All dialysis machines were equipped with AutoSub plus signal analysis software that automatically and continuously adapts the substitution fluid flow according to the blood flow, blood viscosity, and dialyzer characteristics. Percentages of sessions with different types of vascular access were compared regarding achievement of ≥21 L substitution fluid. Logistic regression analysis was conducted to study the independent relationship of selected variables with achievement of ≥21 L substitution volume. Patient- and dialysis-related variables that showed an association with the convection volume were entered in a multivariable model that included hematocrit up front. Respectively, 87%, 84%, and 33% of routine sessions conducted with fistulas, grafts, and catheters qualified as high-volume HDF. Serum albumin levels ≥4.2 g/dL were positively associated with the achievement of at least 21 L substitution volume. Positive associations were also observed for blood flows in the ranges 350-399 and ≥400 mL/min compared with the reference range (300-350 mL/min), for longer treatment time, for fistula versus catheter, for higher filtration fraction, and for dialysis conducted at the end of the week versus Monday. It can be concluded that implementation and sustainability of high-volume HDF is possible in routine clinical practice for almost all patients treated with fistulas and grafts.


Asunto(s)
Hemodiafiltración/métodos , Anciano , Velocidad del Flujo Sanguíneo , Estudios de Cohortes , Femenino , Hematócrito , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Albúmina Sérica/análisis
6.
Biochim Biophys Acta ; 1828(9): 2152-63, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23747364

RESUMEN

Submillimolar concentrations of cytotoxic bile acids (BAs) induce cell death via apoptosis. On the other hand, several cytoprotective BAs were shown to prevent apoptosis in the same concentration range. Still, the mechanisms by which BAs trigger these opposite signaling effects remain unclear. This study was aimed to determine if cytotoxic and cytoprotective BAs, at physiologically active concentrations, are able to modulate the biophysical properties of lipid membranes, potentially translating into changes in the apoptotic threshold of cells. Binding of BAs to membranes was assessed through the variation of fluorescence parameters of suitable derivatized BAs. These derivatives partitioned with higher affinity to liquid disordered than to the cholesterol-enriched liquid ordered domains. Unlabeled BAs were also shown to have a superficial location upon interaction with the lipid membrane. Additionally, the interaction of cytotoxic BAs with membranes resulted in membrane expansion, as concluded from FRET data. Moreover, it was shown that cytotoxic BAs were able to significantly disrupt the ordering of the membrane by cholesterol at physiologically active concentrations of the BA, an effect not associated with cholesterol removal. On the other hand, cytoprotective bile acids had no effect on membrane properties. It was concluded that, given the observed effects on membrane rigidity, the apoptotic activity of cytotoxic BAs could be potentially associated with changes in plasma membrane organization (e.g. modulation of lipid domains) or with an increase in mitochondrial membrane affinity for apoptotic proteins.


Asunto(s)
Ácido Desoxicólico/química , Membrana Dobles de Lípidos/química , Ácido Tauroquenodesoxicólico/química , Ácido Ursodesoxicólico/química , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , Colesterol/química , Difenilhexatrieno , Transferencia Resonante de Energía de Fluorescencia , Colorantes Fluorescentes , Fosfatidilcolinas/química , Fosfatidiletanolaminas/química , Coloración y Etiquetado
7.
Redox Biol ; 58: 102527, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36335761

RESUMEN

Paracrine superoxide (O2•-) and hydrogen peroxide (H2O2) signaling critically depends on these substances' concentrations, half-lives and transport ranges in extracellular media. Here we estimated these parameters for the lumen of human capillaries, arterioles and arteries using reaction-diffusion-advection models. These models considered O2•- and H2O2 production by endothelial cells and uptake by erythrocytes and endothelial cells, O2•- dismutation, O2•- and H2O2 diffusion and advection by the blood flow. Results show that in this environment O2•- and H2O2 have half-lives <60. ms and <40. ms, respectively, the former determined by the plasma SOD3 activity, the latter by clearance by endothelial cells and erythrocytes. H2O2 concentrations do not exceed the 10 nM scale. Maximal O2•- concentrations near vessel walls exceed H2O2's several-fold when the latter results solely from O2•- dismutation. Cytosolic dismutation of inflowing O2•- may thus significantly contribute to H2O2 delivery to cells. O2•- concentrations near vessel walls decay to 50% of maximum 12 µm downstream from O2•- production sites. H2O2 concentrations in capillaries decay to 50% of maximum 22 µm (6.0 µm) downstream from O2•- (H2O2) production sites. Near arterioles' (arteries') walls, they decay by 50% within 6.0 µm (4. µm) of H2O2 production sites. However, they reach maximal values 50 µm (24 µm) downstream from O2•- production sites and decrease by 50% over 650 µm (500 µm). Arterial/olar endothelial cells might thus signal over a mm downstream through O2•--derived H2O2, though this requires nM-sensitive H2O2 transduction mechanisms.


Asunto(s)
Peróxido de Hidrógeno , Superóxidos , Humanos , Células Endoteliales , Cinética , Citosol
8.
Conserv Physiol ; 10(1): coac048, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35875680

RESUMEN

A mechanistic model based on Dynamic Energy Budget (DEB) theory was developed to predict the combined effects of ocean warming, acidification and decreased food availability on growth and reproduction of three commercially important marine fish species: white seabream (Diplodus sargus), zebra seabream (Diplodus cervinus) and Senegalese sole (Solea senegalensis). Model simulations used a parameter set for each species, estimated by the Add-my-Pet method using data from laboratory experiments complemented with bibliographic sources. An acidification stress factor was added as a modifier of the somatic maintenance costs and estimated for each species to quantify the effect of a decrease in pH from 8.0 to 7.4 (white seabream) or 7.7 (zebra seabream and Senegalese sole). The model was used to project total length of individuals along their usual lifespan and number of eggs produced by an adult individual within one year, under different climate change scenarios for the end of the 21st century. For the Intergovernmental Panel on Climate Change SSP5-8.5, ocean warming led to higher growth rates during the first years of development, as well as an increase of 32-34% in egg production, for the three species. Ocean acidification contributed to reduced growth for white seabream and Senegalese sole and a small increase for zebra seabream, as well as a decrease in egg production of 48-52% and 14-33% for white seabream and Senegalese sole, respectively, and an increase of 4-5% for zebra seabream. The combined effect of ocean warming and acidification is strongly dependent on the decrease of food availability, which leads to significant reduction in growth and egg production. This is the first study to assess the combined effects of ocean warming and acidification using DEB models on fish, therefore, further research is needed for a better understanding of these climate change-related effects among different taxonomic groups and species.

9.
Clin Exp Nephrol ; 15(1): 159-63, 2011 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21069411

RESUMEN

Atheroembolic renal disease is caused by foreign-body reaction to cholesterol crystals flushed from the atherosclerotic plaques into the small-vessel system of the kidneys. It is an underdiagnosed entity, mostly related to vascular procedures and/or anticoagulation, and prognosis is considered to be poor. Besides the benefit of aggressive medical prevention of further embolic events, use of steroid therapy has been associated with greater survival. Here we report a case of a patient with a multisystemic presentation of the disease days after performance of percutaneous coronary intervention and anticoagulation initiation due to an episode of myocardial infarction. Renal, cutaneous, ophthalmic, neurological, and possibly muscular and mesenteric involvement was diagnosed. Although medical treatment with corticosteroids and avoidance of further anticoagulation was applied, the patient rapidly progressed to end-stage renal disease requiring hemodialysis and died 6 months after diagnosis. This is a case of catastrophic progression of the disease resistant to therapeutic measures. Focus on diagnosis and more efficient preventive and therapeutic protocols are therefore needed.


Asunto(s)
Embolia por Colesterol/complicaciones , Embolia por Colesterol/mortalidad , Reacción a Cuerpo Extraño/complicaciones , Enfermedades Renales/etiología , Enfermedades Renales/mortalidad , Anciano , Embolia por Colesterol/inmunología , Embolia por Colesterol/patología , Resultado Fatal , Humanos , Enfermedades Renales/patología , Enfermedades Renales/fisiopatología , Fallo Renal Crónico/mortalidad , Fallo Renal Crónico/fisiopatología , Masculino , Diálisis Renal
10.
Artículo en Inglés | MEDLINE | ID: mdl-31370308

RESUMEN

Coastal areas have been increasingly affected by human activities, marine pollution and climate change are among the most important pressures affecting these environments. Human-induced pressures occur in a cumulative way and generate additive, antagonistic or synergistic effects. Knowledge on synergistic effects is crucial to coastal zone management, since they may imply a change in human uses of these systems, as well as dedicated action plans in order to reduce hazards and environmental risks. In this work, we provide an overview of the available literature on synergistic effects between climate change and chemical pollution, and discuss current knowledge, methodological approaches, and research gaps and needs. Interactions between these two pressures may be climate change dominant (climate change leads to an increase in contaminant exposure or toxicity) or contaminant-dominant (chemical exposure leads to an increase in climate change susceptibility), but the mechanistic drivers of such processes are not well known. Results from a few meta-analyses studies and reviews showed that synergistic interactions tend to be more frequent compared to additive and antagonistic ones. However, most of the studies are individual-based and assess the cumulative effects of a few contaminants individually in laboratory settings together with few climate variables, particularly temperature and pH. Nevertheless, a wide diversity of contaminants have already been individually tested, spanning from metals, persistent organic pollutants and, more recently, emergent pollutants. Population and community based approaches are less frequent but have generated very interesting and more holistic perspectives. Methodological approaches are quite diverse, from laboratory studies to mesocosm and field studies, or based on statistical or modelling tools, each with their own potential and limitations. More holistic comparisons integrating several pressures and their combinations and a multitude of habitats, taxa, life-stages, among others, are needed, as well as insights from meta-analyses and systematic reviews.


Asunto(s)
Cambio Climático , Ecosistema , Océanos y Mares , Contaminación Química del Agua/efectos adversos , Actividades Humanas , Humanos
11.
Methods Mol Biol ; 1981: 99-115, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31016650

RESUMEN

Cholestasis is characterized by impaired secretion of bile flow that can result in the accumulation of highly abnormal levels of bile acids causing hepatocyte and biliary injury. As amphipathic molecules, bile acids can intercalate in lipid membranes, and pathophysiologic concentrations of bile acids have the potential to induce marked changes in the biophysical properties of biomembranes, including membrane ordering. These effects, particularly on the mitochondrial and plasma membranes, have been proposed to trigger toxicity of bile acids. This chapter details different fluorescence-based methods (fluorescence polarization, and spectroscopy/imaging of solvatochromic dyes) to evaluate the impact of different bile acids on membrane order. Protocols are described for the application of these methods to biomimetic vesicles, isolated mitochondria, and hepatocytes, following a bottom-up approach.


Asunto(s)
Ácidos y Sales Biliares/metabolismo , Hepatocitos/metabolismo , Mitocondrias/metabolismo , Animales , Ácidos y Sales Biliares/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Polarización de Fluorescencia , Hepatocitos/efectos de los fármacos , Mitocondrias/efectos de los fármacos
12.
Phys Life Rev ; 20: 78-84, 2017 03.
Artículo en Inglés | MEDLINE | ID: mdl-28237359

RESUMEN

In response to the comments on review "Physics of metabolic organization", we discuss the universality and the future prospects of physiological energetics. The topics range from the role of entropy in modeling living organisms to the apparent ubiquity of the von Bertalanffy curve, and the potential applications of the theory in yet unexplored domains. Tradeoffs in outreach to non-specialists are also briefly considered.


Asunto(s)
Entropía , Física , Humanos
13.
Phys Life Rev ; 20: 1-39, 2017 03.
Artículo en Inglés | MEDLINE | ID: mdl-27720138

RESUMEN

We review the most comprehensive metabolic theory of life existing to date. A special focus is given to the thermodynamic roots of this theory and to implications that the laws of physics-such as the conservation of mass and energy-have on all life. Both the theoretical foundations and biological applications are covered. Hitherto, the foundations were more accessible to physicists or mathematicians, and the applications to biologists, causing a dichotomy in what always should have been a single body of work. To bridge the gap between the two aspects of the same theory, we (i) adhere to the theoretical formalism, (ii) try to minimize the amount of information that a reader needs to process, but also (iii) invoke examples from biology to motivate the introduction of new concepts and to justify the assumptions made, and (iv) show how the careful formalism of the general theory enables modular, self-consistent extensions that capture important features of the species and the problem in question. Perhaps the most difficult among the introduced concepts, the utilization (or mobilization) energy flow, is given particular attention in the form of an original and considerably simplified derivation. Specific examples illustrate a range of possible applications-from energy budgets of individual organisms, to population dynamics, to ecotoxicology.


Asunto(s)
Biofisica/métodos , Metabolismo , Animales , Homeostasis , Humanos , Modelos Biológicos
15.
Phys Rev E Stat Nonlin Soft Matter Phys ; 74(5 Pt 1): 051901, 2006 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17279933

RESUMEN

We carry out a thermodynamic analysis to an organism. It is applicable to any type of organism because (1) it is based on a thermodynamic formalism applicable to all open thermodynamic systems and (2) uses a general model to describe the internal structure of the organism--the dynamic energy budget (DEB) model. Our results on the thermodynamics of DEB organisms are the following. (1) Thermodynamic constraints for the following types of organisms: (a) aerobic and exothermic, (b) anaerobic and exothermic, and (c) anaerobic and endothermic; showing that anaerobic organisms have a higher thermodynamic flexibility. (2) A way to compute the changes in the enthalpy and in the entropy of living biomass that accompany changes in growth rate solving the problem of evaluating the thermodynamic properties of biomass as a function of the amount of reserves. (3) Two expressions for Thornton's coefficient that explain its experimental variability and theoretically underpin its use in metabolic studies. (4) A mechanism that organisms in non-steady-state use to rid themselves of internal entropy production: "dilution of entropy production by growth." To demonstrate the practical applicability of DEB theory to quantify thermodynamic changes in organisms we use published data on Klebsiella aerogenes growing aerobically in a continuous culture. We obtain different values for molar entropies of the reserve and the structure of Klebsiella aerogenes proving that the reserve density concept of DEB theory is essential in discussions concerning (a) the relationship between organization and entropy and (b) the mechanism of storing entropy in new biomass. Additionally, our results suggest that the entropy of dead biomass is significantly different from the entropy of living biomass.


Asunto(s)
Ingestión de Alimentos/fisiología , Ingestión de Energía/fisiología , Metabolismo Energético/fisiología , Enterobacter aerogenes/crecimiento & desarrollo , Crecimiento/fisiología , Modelos Biológicos , Dinámica Poblacional , Animales , Proliferación Celular , Simulación por Computador , Entropía , Humanos , Termodinámica
16.
Chemosphere ; 146: 330-7, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26735734

RESUMEN

Ochrobactrum tritici SCII24T bacteria is an environmental strain with high capacity to resist to arsenic (As) toxicity, which makes it able to grow in the presence of As(III). The inactivation of the two functional arsenite efflux pumps, ArsB and ACR3_1, resulted in the mutant O. tritici As5 exhibiting a high accumulation of arsenite. This work describes a method for the immobilization of the mutant cells O. tritici As5, on a commercial polymeric net after sputtered modified by the deposition of poly(tetrafluoroethylene) (PTFE) thin films, and demonstrates the capacity of immobilized cells to accumulate arsenic from solutions. Six different set of deposition parameters for PTFE thin films were developed and tested in vitro regarding their ability to immobilize the bacterial cells. The surface that exhibited a mild zeta potential value, hydrophobic characteristics, the lowest surface free energy but with a high polar component and the appropriate ratio of chemical reactive groups allowed cells to proliferate and to grow as a biofilm. These immobilized cells maintained their ability to accumulate the surrounding arsenite, making it a great arsenic biofilter to be used in bioremediation processes.


Asunto(s)
Arsenitos/metabolismo , Ochrobactrum/metabolismo , Politetrafluoroetileno/metabolismo , Eliminación de Residuos Líquidos/métodos , Contaminantes Químicos del Agua/metabolismo , Polímeros
17.
PLoS One ; 10(7): e0131317, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26132104

RESUMEN

Ochrobactrum tritici SCII24T is a highly As-resistant bacterium, with two previously described arsenic resistance operons, ars1 and ars2. Among a large number of genes, these operons contain the arsB and Acr3 genes that encode the arsenite efflux pumps responsible for arsenic resistance. Exploring the genome of O. tritici SCII24T, an additional putative operon (ars3) was identified and revealed the presence of the Acr3_2 gene that encodes for an arsenite efflux protein but which came to prove to not be required for full As resistance. The genes encoding for arsenite efflux pumps, identified in this strain, were inactivated to develop microbial accumulators of arsenic as new tools for bioremediation. Six different mutants were produced, studied and three were more useful as biotools. O. tritici wild type and the Acr3-mutants showed the highest resistance to As(III), being able to grow up to 50 mM of arsenite. On the other hand, arsB-mutants were not able to grow at concentrations higher than 1 mM As(III), and were the most As(III) sensitive mutants. In the presence of 1 mM As(III), the strain with arsB and Acr3_1 mutated showed the highest intracellular arsenic concentration (up to 17 ng(As)/mg protein), while in assays with 5 mM As(III), the single arsB-mutant was able to accumulate the highest concentration of arsenic (up to 10 ng(As)/mg protein). Therefore, arsB is the main gene responsible for arsenite resistance in O. tritici. However, both genes arsB and Acr3_1 play a crucial role in the resistance mechanism, depending on the arsenite concentration in the medium. In conclusion, at moderate arsenite concentrations, the double arsB- and Acr3_1-mutant exhibited a great ability to accumulate arsenite and can be seen as a promising bioremediation tool for environmental arsenic detoxification.


Asunto(s)
Arsénico/toxicidad , ATPasas Transportadoras de Arsenitos/genética , Proteínas Bacterianas/genética , Contaminantes Ambientales/toxicidad , Regulación Bacteriana de la Expresión Génica , Mutación , Ochrobactrum/efectos de los fármacos , ATPasas Transportadoras de Arsenitos/deficiencia , Proteínas Bacterianas/metabolismo , Biodegradación Ambiental , Farmacorresistencia Bacteriana/genética , Ingeniería Genética , Transporte Iónico , Ochrobactrum/genética , Ochrobactrum/metabolismo , Operón
18.
Metallomics ; 5(4): 390-7, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23487302

RESUMEN

Severe environmental problems arise from old uranium mines, which continue to discharge uranium (U) via acid mine drainage water, resulting in soil, subsoil and groundwater contamination. Bioremediation of U contaminated environments has been attempted, but most of the conceptual models propose U removal by cell suspensions of anaerobic bacteria. In this study, strain Rhodanobacter A2-61, isolated from Urgeiriça Mine, Portugal, was shown to resist up to 2 mM of U(vi). The conditions used (low nutrient content and pH 5) potentiated the interaction of the toxic uranyl ion with the tested strain. The strain was able to remove approximately 120 µM of U(vi) when grown aerobically in the presence of 500 µM U. Under these conditions, this strain was also able to lower the phosphate concentration in the medium and increased its capacity to take up inorganic phosphate, accumulating up to 0.52 µmol phosphate per optical density unit of the medium at 600 nm, after 24 hours, corresponding approximately to the late log phase of the bacterial culture. Microscopically dense intracellular structures with nanometer size were visible. The extent of U inside the cells was quantified by LS counting. EDS analysis of heated cells showed the presence of complexes composed of phosphate and uranium, suggesting the simultaneous precipitation of U and phosphate within the cells. XRD analysis of the cells containing the U-phosphate complexes suggested the presence of a meta-autunite-like mineral structure. SEM identified, in pyrolyzed cells, crystalline nanoparticles with shape in the tetragonal system characteristic of the meta-autunite-like mineral structures. U removal has been reported previously but mainly by cell suspensions and through release of phosphate. The innovative Rhodanobacter A2-61 can actively grow aerobically, in the presence of U, and can efficiently remove U(vi) from the environment, accumulating it in a structural form consistent with that of the mineral meta-autunite inside the cell, corresponding to effective metal immobilization. This work supports previous findings that U bioremediation could be achieved via the biomineralization of U(vi) in phosphate minerals.


Asunto(s)
Espacio Intracelular/metabolismo , Fosfatos/metabolismo , Uranio/metabolismo , Xanthomonadaceae/metabolismo , Aerobiosis/efectos de los fármacos , Biodegradación Ambiental/efectos de los fármacos , Espacio Intracelular/efectos de los fármacos , Iones , Fosfatos/farmacología , Polvos , Espectrometría por Rayos X , Uranio/aislamiento & purificación , Difracción de Rayos X , Xanthomonadaceae/efectos de los fármacos , Xanthomonadaceae/crecimiento & desarrollo , Xanthomonadaceae/ultraestructura
19.
Clin Kidney J ; 5(4): 352-5, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25874097

RESUMEN

In alkaptonuria, deficiency of homogentisate 1,2-dioxygenase leads to the accumulation of homogentisic acid (HGA) and its metabolites in the body, resulting in ochronosis. Reports of patients with alkaptonuria who have decreased kidney function are rare, but this seems to play an important role in the natural history of the disease. We describe a 68-year-old female with chronic kidney disease (CKD) of unknown etiology who started peritoneal dialysis (PD) after 5 years of follow-up and who was diagnosed with alkaptonuria at this time. Progressive exacerbation of ochronotic manifestations had been noted during these last few years, as kidney function worsened. After PD initiation, the disease continued to progress, and death occurred after one year and a half, due to severe aortic stenosis-related complications. Her 70-year-old sister was evaluated and also diagnosed with alkaptonuria. She had no renal dysfunction. Higher HGA excretion and significantly milder ochronosis than that of her sister were found. We present two alkaptonuric sisters with similar comorbidities except for the presence of CKD, who turned out to have totally different evolutions of their disease. This report confirms that kidney dysfunction may be an important factor in determining the natural history of alkaptonuria.

20.
Cad. saúde colet., (Rio J.) ; 25(3): 324-331, jul.-set. 2017. tab
Artículo en Portugués | LILACS | ID: biblio-890032

RESUMEN

Resumo Introdução A mortalidade materna representa um evento de grande magnitude no Brasil e no mundo, refletindo a falta de acesso aos serviços de saúde e as condições socioeconômicas desfavoráveis. Objetivo Relacionar as alterações maternas com o desfecho gravídico-puerperal no óbito materno. Métodos Estudo transversal e retrospectivo, conduzido com dados coletados de 53 prontuários de gestantes e puérperas que foram a óbito em um hospital estadual referência em gestação de alto risco em Goiás. Resultados A mortalidade ocorreu mais frequentemente em mulheres com idade entre 20 e 34 anos (76%), solteiras (55%), primigestas (38%), procedentes do interior do Estado (60%) e com parto na instituição do estudo (62%). As principais causas obstétricas de óbito foram as diretas (55%), com predomínio no período puerperal (83%). A razão da mortalidade materna correspondeu a 228,4 por 100 mil nascidos vivos. Houve uma associação significativa entre as complicações no parto e o local de ocorrência do parto (P=0,001). Conclusão O óbito materno ocorreu em solteiras, com idade reprodutiva, procedentes de outros municípios, por causas diretas e no puerpério. Ressalta-se a necessidade de melhorar o acesso aos serviços de saúde em obstetrícia, pois a maioria dos casos pode ser prevenida.


Abstract Introduction Maternal mortality represents an event of great magnitude in Brazil and worldwide. The occurrence of maternal death reflects the lack of access to health services and unfavorable socioeconomic conditions. Objective To correlate maternal changes and puerperal pregnancy outcome in maternal death. Methods Cross-sectional and retrospective study. We used data collected from 53 medical records of pregnant women and postpartum women who died at a Reference State Hospital in high risk gestation in Goiás. Results Mortality occurred in women aged 20 to 34 years (76%), single (55%), first pregnancy (38%), from the interior of the State (60%) and 62% had the childbirth at the institution of the study. The main obstetric causes were 55% direct and 24% indirect, being predominant in the puerperal period (83%). The maternal mortality ratio was 228.4. There was a significant association between complications at childbirth and the place of birth (P = 0.001). Conclusion The maternal death occurred in single women, of reproductive age, coming from other counties, for direct causes and in the puerperium. It is necessary to improve the access to health services in obstetrics, since most cases are preventable.

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